Project Summary The Lung Phenotyping Core, designated as Core C, is a centralized facility that provides primary murine alveolar epithelial cells and macrophages from lungs, as well as satellite cells and muscle to each of the three projects and Core B. Core C also maintains secondary lung cell lines for the PPG. Centralization of the cell culture facilities will ensure that continuous supply of high-quality primary alveolar epithelial cells are available to each of the three projects and Core B. Core C will use advanced flow cytometry capabilities to quantify, phenotype and sort specific inflammatory and parenchymal cell populations from the murine lung. The sorting and isolation capabilities of the Core will allow cell type specific assessment of proteostasis networks via mass spectroscopy (as outlined by Core B). Core C will also isolate muscle satellite cells from mouse skeletal muscles; we have established the optimal culture conditions for cell proliferation and myotube formation of mouse satellite cells. The Core personnel have extensive experience in the isolation of primary alveolar epithelial cells and macrophages from human and mouse lungs, as well as general cell culture techniques. The consolidated cell culture facility provides an economical means of isolating and culturing primary and secondary cells. This translates into reduced overall costs (i.e., personnel, reagents, and animals) and more importantly maintains the quality of the cells used in research. The Core will also provide investigators with well-characterized mouse models of influenza A-induced pneumonia and quantitative measurements of lung inflammation, lung injury, and muscle function will be conducted by Core C according to the research plan outlined in Projects 1, 2, and 3. In addition, Core C will conduct RNA analysis to assess the relationship between specific gene expression and proteostasis using the Northwestern University Next-Generation Sequencing (NGS) Core Facility. High throughput sequencing technology will be used to study transcriptome dynamics. Additionally, the project investigators have proposed several murine strains, many of which will be bred with cell-type or tissue specific Cre recombinase lines to induce a tissue-specific knockout by Core C. Many of the strains will be used in more than one project, highlighting the synergy and economies of scale achievable by this Program Project.